Examining the role of IL18R1 and IL18RAP in asthma

About the Project

Background

IL-18 is a proinflammatory cytokine though to be involved in the induction and regulation of Type 1 immune responses (1). It functions by binding to the IL18 receptor which is composed of two proteins, IL-18 receptor 1 (IL18R1) and IL-18 receptor accessory protein (IL18RAP) (2). Both genes are found in a gene cluster on human chromosome 2q that has been associated with asthma in genome wide association studies (3). The precise role of IL18 and its receptor in asthma pathogenesis is unclear.

Aims

To investigate expression and function of the IL-18 receptor on immune cell types in peripheral blood samples from asthma and control subjects

Objectives

• Clone IL18R1 and IL18RAP genes into a lentiviral expression vector containing a fluorescent reporter
• Overexpress IL18R1 and IL18RAP in HEK293 cells
• Test commercially available antibodies recognizing IL18R1 and IL18RAP by flow cytometry and western blot.
• Examine expression of IL18R1 on immune cell types in peripheral blood samples.
• Using in vitro assays to examine the effect of IL18 on immune cell function.
• Compare effect of IL18 on immune cell function in healthy and asthmatic subjects.

Experimental plan and techniques to be used

• Molecular Biology including restriction digestion, PCR, DNA sequencing, transfection of mammalian cells
• Lentiviral particle production and transduction of primary human immune cells
• Advanced multi colour flow cytometry
• Real-time quantitative RT-PCR using TaqMan chemistry
• Western blotting

Apply at: https://le.ac.uk/study/research-degrees/research-subjects/respiratory-sciences

PhD entry requirements: https://le.ac.uk/study/research-degrees/entry-reqs

Supervisor contact details:

Prof David Cousins - dc282@leicester.ac.uk